This protocol was developed to detect TSWV in potato leaves
and tubers. It has also been used to detect TSWV
in its thrips vector insects. The details of the protocol are
not reported here, as this test is being further developed by
Agdia, Inc.
Experimental Approach - TSWV
Detection
1. Extract RNA from samples:
Positive controls (TSWV-inoculated plants)
Negative controls (Tissue
culture plantlets)
Unknowns
(symptomatic tissue, shown above)
2. Synthesize cDNA using DNA oligo primers
homologous to viral sequence conserved among TSWV isolates
3. Perform
PCR amplification of cDNA fragment using DNA oligo primers homologous to viral sequence conserved
among TSWV isolates
Interpreting Results - TSWV Detection
1. Analyze a portion of the PCR products by agarose gel electrophoresis
2. Compare product of
"unknown" reaction with the positive and negative controls
3. Product
accumulation in "unknown" reaction is a positive result
4. Products of
a positive test can be further analyzed by sequencing to identify isolate
present in sample
Mail inquiries to TLG@entomology.wisc.edu
Updated 11Jan2007.